This renewal application proposes to complete the development of enhanced serological and virological test systems for B virus (Cercopithecine herpesvirus 1, Herpesvirus simiae) as well as other simian herpesviruses, to unambiguously identify B virus infection in humans, macaques, or other exposed nonhuman primates. The applicants will use structural and functional properties of B virus and closely related herpesvirus to complete development of a multifaceted diagnostic system with the following assay systems: (a) a rapid enzyme-immunoassay for serological identification of B virus propagation; (b) a rapid antigen detection system to identify the presence of replication competent B virus present in human and nonhuman primate tissues (lesional or nonlesional sites using monoclonal antibodies; and (c) molecular analysis using in situ hybridization (ISH) coupled with polymerase chain reaction (PCR) techniques to identify primates with potential for reactivation of latent B virus. The applicants will accomplish the objective with the following Specific Aims: (1) identify glycoproteins and proteins from which each of the infected cell polypeptides/glycopeptides are derived; (2) sequence major gene homologues encoded by B virus and identify divergent and conserved regions with respect to function; (3) express selected sequences or domains to produce recombinant determinants of B virus polypeptides/glycopeptides; (4) utilize monoclonal and polyclonal monospecific sera prepared against synthetic peptides and recombinant fusion proteins for rapid assays to identify selected antibody and antigen specificities; (5) identify human monoclonal antibody specificities from infection survivors versus individuals who have succumbed to infection; and (6) identify and quantify the efficacy of novel antiviral agents that can be made available in the event of new or reactivated zoonotic infection with B virus. The results of the research proposed in this renewal application will provide the basis for implementing the safest possible operating standards for NIH and other federally supported macaque resource colonies and their personnel. Improved technologies and reagents will enhance management, prevention, and control of B virus infections in humans and other primate species. Development of these enhanced and unique diagnostic systems is especially important to prevent further zoonotic disease in humans and insure continued usage of macaques for AIDS research.